Oligonucleotide Synthesis Devices

Orochem OF1100TM Plates

The OF1100 96 Well filter plate is manufactured exclusively by Orochem Technologies. The Polypropylene filter plate has 0.7ml/well volume and comes fritted with a 10um polyethylene frit.

Sold worldwide for applications in Oligonucleotide synthesis, solid phase extraction, Filtration and clean up of PCR products prior to sequencing.

  • Features

    • Consistent Quality
    • Fully Integrated Manufacturing
    • High Throughput
    • Excellent Flowrate

High Throughput DNA Synthesis in 96 Well Plates
Now available Pre-dispensed with Universal Support

GENOMICS OLIGONUCLEOTIDE SYNTHESIS DEVICES

Orochem 96 Well Collection Plates

  • Also available is the 1.0mL OT-850353 deep well collection plate compatible with Mermade (Bioautomation, Plano, TX) and PolyPlex (Gene Machines) equipment for solid phase synthesis.

Individual Deoxynucleoside CPG Solid Support Columns

In conjunction with the OF1100, Orochem offers state of the art CPG support for DNA and RNA synthesis.
Orochem CPG supports for Oligo Synthesis have the following features:

  • Uses Standard Phosphoramidites: dA-Bz, dG-dmf, dC-Ac and T, dGIBu, dC-Bz
  • Standard Cycle Time/ Program
  • Easily scalable approach: 50 nM-1μM
  • Big cost savings
  • Adaptable to automation

Oligonucleotide Deprotection using Orochem’s Proprietary Non- Aqueous Cocktail

Orochem’s Novel Synthesis Procedure Using Non-Aqueous Cocktail

  • Cleavage and De-protection treatment (500μl) at room temperature for 45 minutes in the synthesis column (ABI-3900 or 394) or Orochem’s CPG cartridges
  • Washing with organic solvent (500μl).
  • Dry with air and Elute in water or TE buffer (250μl). Ready for use.

Oligonucleotide Purification

Oligonucleotide Purification

Orochem has the following purification options available to remove the impurities generated during the synthesis of modified oligonucleotides. Which purification method is employed depends on how the oligonucleotide is modified and how it will be used.

1. A) Desalting (Oligo using OrpheusTM Silica C18 cartridges)
Desalting procedures remove inorganic salts, traces of organic compounds, low molecular weight impurities, and short failure sequences. Desalting is accomplished by taking advantage of size (molecular weight) or solubility differences.

1. B) High Throughput Desalting with C18 in 96 Well Filter Plates

GENOMICS OLIGONUCLEOTIDE PURIFICATION

2. Gel Filtration (Desalting Oligos using Sephadex Packed Cartridges & Plates)
Gel filtration removes lower molecular weight molecules, inorganic salts and very short failure sequences. Purification occurs by passing a sample through Sephadex column, which separates molecules based on size.

3.Reverse Phase cartridge Purification Using OPC OroPure Cartridges
These cartridges work on the principle of adsorption the dimethyltrityl (DMT) group of the trityl-on oligonucleotide while non-DMT bearing failure sequences, by-products, and other impurities wash through. The DMT group is removed with mild acid, allowing the purified, detritylated oligonucleotide to be eluted. This cartridge not only eliminates salts and trace organic impurities from the crude oligonucleotide, but also provides pure oligonucleotides for sequencing primers, PCR primers, hybridization probes, and gene constructions

GENOMICS OLIGONUCLEOTIDE PURIFICATION
GENOMICS OLIGONUCLEOTIDE PURIFICATION CHART

Demonstration of Oligo Synthesis with Universal Support Packed into OF1100 and Synthesized on the Biolytic 192 Synthesizers
Characteristics of the Oligos Obtained with the New Orochem’s Method

  • Purity >92% by HPLC and PAGE analysis
  • Biological activity: PCR, Gene Assembly
  • TOTAL TIME: 2 hours
  • NB: No drying. No de-salting

Analytical Data: HPLC

Analytical Data: PAGE Analysis

RELIASIL LC SOLUTIONS
OROCHEM Reliasil HPLC Columns for DNA/RNA, Proteins, and Peptides Purification
Oligo-Reliasil columns were designed by Orochem Technologies Inc. specifically for Reversed Phase Liquid Chromatography. A proprietary type A silica manufacturing process which yields a Gaussian particle distribution, narrow pore-size distribution, and strong resistance to high pH is used. The resulting material is perfect for packing high-efficiency columns designed for purification and characterization purposes.
Advantages & Features:

  • High Resolution with maximum purity
  • Various spherical sizes- 3, 5 & 10μm
  • High Efficiency Pore size- 300A
  • Advanced surface chemistries: C8, C18
  • Stable chemistry bonded phases to avoid non-specific binding
  • Ideal scale-up production from analytical to preparative mode
  • Affordable lower cost with standard micro bore analytical hardware.
Material CharacteristicRELIASIL
 Reversed PhaseIon Exchange
Particle SupportSilicaHigh Purity Silica
Bonded PhaseC18SAX
Particle Size3, 5 & 10 μm3, 5 & 10 μm
Pore Size300A300A
Surface Area190190
pH Stability1.0-12.01.5-11.0
pH Effective Range2.0-9.54.0-8.5

DNA Purification on RELIASIL Columns

HPLC Conditions
Column: Reliasil C18, 3μ, 300A
Dimensions: 50 x 4.6mm
Part No: 21001014
Mobile Phase
A: 50mM TEAA, pH 7.5/ 5% Acetonitrile
B: 100% Methanol
Gradient: 0 to 80% B in 20 minutes
Flow Rate: 1mL/min
Detection: UV@260nm
Sample: 40 nucleotide DNA sequence
Load: 20 μL (2.5 μg of DNA)
HPLC Conditions
Column: Reliasil C18, 3μ, 300A
Dimensions: 50 x 4.6mm
Part No: 21001014
Mobile Phase
A: 50mM TEAA, pH 7.5/ 5% Acetonitrile
B: 100% Methanol
Gradient: 0 to 20% B in 20 minutes
Flow Rate: 1mL/min
Detection: UV@260nm
Sample: Biotinylated 40nt-DNA sequence
Load: 20 μL (5 μg of DNA)
HPLC Conditions
Column: Reliasil C18, 5μ, 300A
Dimensions: 240 x 4.6mm
Part No: R5HI-126
Mobile Phase
A: 100mM TEAA
B: 100mM TEAA, pH 7.5 in Acetonitrile
Gradient: 0 to 30% B in 30 minutes
Flow Rate: 1mL/min
Detection: UV@260nm
Sample: NH2 labeled DNA sample
Load: 20 μL (5 μg of DNA)